EP0113215A2 - Microorganisms of the species Lactobacillus italicus, processes for their preparation and their use in the production of lactic acid - Google Patents

Microorganisms of the species Lactobacillus italicus, processes for their preparation and their use in the production of lactic acid Download PDF

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Publication number
EP0113215A2
EP0113215A2 EP83307542A EP83307542A EP0113215A2 EP 0113215 A2 EP0113215 A2 EP 0113215A2 EP 83307542 A EP83307542 A EP 83307542A EP 83307542 A EP83307542 A EP 83307542A EP 0113215 A2 EP0113215 A2 EP 0113215A2
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growth
acid
microorganism
italicus
negative
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EP0113215B1 (en
EP0113215A3 (en
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Takayama Kenichiro
Masunaga Isumi
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KH Neochem Co Ltd
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Kyowa Hakko Kogyo Co Ltd
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    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12PFERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P7/00Preparation of oxygen-containing organic compounds
    • C12P7/40Preparation of oxygen-containing organic compounds containing a carboxyl group including Peroxycarboxylic acids
    • C12P7/56Lactic acid
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20Bacteria; Culture media therefor
    • C12N1/205Bacterial isolates
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12RINDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00Microorganisms ; Processes using microorganisms
    • C12R2001/01Bacteria or Actinomycetales ; using bacteria or Actinomycetales
    • C12R2001/225Lactobacillus

Definitions

  • the present invention relates to microorganisms of the species Lactobacillus italicus, processes for their preparation and their use in the production of lactic acid.
  • the lactobacilli may be isolated from a dough for making bread.
  • Lactobacillus sanfrancisco is a known microorganism isolated from a dough for sour dough French bread conventionally made in the San Francisco Bay area of the United States, and is useful in the preparation of a liquid starter and dough for sour dough bakery products such as sour dough French bread (U.S. Patents Nos. 3,734,743 and 3,891,773).
  • This microorganism is capable of growing in the presence of a sour dough yeast, viz
  • microorganisms from a dough used for the preparation of panettone which is a type of Italian rich bread.
  • the present invention is based upon the discovery that the microorganisms which we have isolated are capable of growing in the absence of yeast and are of potential interest as lactic acid-producing microorganisms.
  • microorganisms in one embodiment may be further characterised in that glucose and maltose are the sole assimilable carbon sources.
  • microorganisms of the present invention have been designated by us as Lactobacillus italicus for reasons hereinafter described and include the following two strains:
  • the present strain appears as rods having a size of (0.8-1.0) x (2 -7) ⁇ , and occurs in the form of a single rod or in chains.
  • This strain is non-motile, non- sporulating and Gram-positive.
  • composition of ML medium Composition of ML medium:
  • this strain is cultured on an ML agar plate medium prepared by adding 20 g of agar to ML medium at 30°C for 2 days, round, smooth, entire, raised, semi-transparent and milky white colonies are formed, the diameter of the colonies being not greater than about 1 mm.
  • the culture exhibits a moderate turbidity. No growth on the surface layer. Flocculent sediment is formed.
  • An unstaturated fatty acid is not required for growth on an ML medium.
  • Maltose and glucose are assimilable, although maltose is more strongly assimilated than glucose.
  • L. italicus TL-1 and L. italicus SH-150 are Gram-positive rods which are non- sporulating, non-motile, anaerobic to facultative aerobic and catalase negative. Moreoever, they are capable of producing lactic acid and ethyl alcohol from glucose and maltose. Thus, it is apparent that they are classified into the genus Lactobacillus. However, the present strains differ from all lactobacilli disclosed, for example, in Bergey's Manual of Determinative Bacteriology, 8th Edition (1974) in so far as the following identifying characteristics are concerned:-
  • microorganisms of the present invention designated by us as Lactobacillus italicus are new microorganisms.
  • a process for the preparation of a microorganism as hereinbefore defined which comprises inoculating an agar medium with an aqueous suspension of panettone dough, incubating the inoculated medium under anaerobic conditions to form colonies on said medium, isolating Lactobacillus italicus having the identifying characteristics hereinbefore defined and culturing the isolated strain(s) in or on a sterile medium.
  • Panettone dough is described in Bakers Digest October 76 80 (1977).
  • L. italicus of the present invention may be cultured, for example, in the following manner:-
  • Various solid and liquid media may be used for culturing L. italicus in so far as they are suitable for culturing lactobacilli and contain glucose and maltose as carbon sources.
  • carbon sources various naturally occurring substances containing glucose and maltose such as, for example, starch hydrolyzate liquor, wheat flour and the like may.beused.
  • various organic acids such as, for example, acetic acid, citric acid and the like may be added to the medium in order to promote the growth of L. italicus.
  • various materials containing organic nitrogen such as, for example, yeast extract, peptone, meat extract, casein, corn steep liquor, tomato juice and the like as nitrogen sources is advantageous for promoting the growth of L.
  • the medium inorganic compounds such as, for example, ammonium sulfate, ammonium nitrate, ammonium phosphate and the like. It is also advantageous to add to the medium, for example, phosphoric acid, various ions of metals such as e.g. potassium, sodium, magnesium, manganese, iron, molybdenum and the like.
  • phosphoric acid various ions of metals such as e.g. potassium, sodium, magnesium, manganese, iron, molybdenum and the like.
  • cysteine, cystine, glutathione, ascorbic acid and the like is also advantageous. in order to reduce the partial pressure of oxygen.
  • an unsaturated fatty acid is required for the growth of the cultured microorganism, it is necessary to add a suitable agent such as, for example, oleic acid, Tween 80 (a surface active agent, commercially available from Atlas Chemical Industries, Inc., U.S.A.), lectin and the like to the medium.
  • a suitable agent such as, for example, oleic acid, Tween 80 (a surface active agent, commercially available from Atlas Chemical Industries, Inc., U.S.A.), lectin and the like to the medium.
  • the culturing may for example be effected at a temperature of from 15 to 38°C, preferably from 28 to 32°C and at a pH of from 4.2 - 6.5,especially from 5.4 to 5.8 for 1 to 2 days.
  • the culturing may preferably be effected under anaerobic conditions in the presence of carbon dioxide.
  • the mixing ratio of carbon dioxide with air may be from 25 to 95 % by volume.
  • ML medium as hereinbefore defined in order to isolate the present strain from a dough used for the preparation of panettone as hereinbefore described.
  • a panettone dough is suspended in a sterilized physiological saline solution, suitable buffer solution and the like and the mixture is smeared, for example, on an ML agar plate medium as hereinbefore defined for culturing at a temperature of from 28 to 32°C for 1-2 days under anaerobic conditions to obtain a pure culture of L. italicus of the present invention.
  • a process for the preparation of lactic acid which comprises culturing a microorganism as hereinbefore defined in or on a medium therefor whereby to obtain a cultured liquor containing lactic acid.
  • the lactic acid obtained is preferably isolated from the obtained cultured liquor.
  • a panettone dough purchased from the Italian market (0.3 g) was suspended in a physiological saline solution (10 ml) under. sterilized conditions. The suspension thus-prepared was multiply diluted with the same solution. Then, on each occasion, the diluted dough suspension (0.1 ml) was smeared on an ML medium as hereinbefore defined, and a Gas Pak System (Commercial Product of Baltimore Biological Laboratories, Inc., U.S.A.) was used to provide an atmosphere of carbon dioxide, in which the culturing was effected at a temperature of 30°C for 2 days to obtain semi-transparent, milky white colonies having a diameter of not greater than about 1 mm.
  • a Gas Pak System Common Product of Baltimore Biological Laboratories, Inc., U.S.A.
  • the thus-prepared medium was put into a large test tube and was sterilized at a temperature of 120°C for 15 minutes in an autoclave.
  • Each medium was then inoculated with L. italicus TL-1 (FERM-BP 189) or L. italicus SH-150 (FERM-BP 190) (about 10 8 cells per ml) obtained from the ML agar plate medium hereinbefore described and incubated at a temperature of 30°C for the period shown in Table 1.
  • the resultant growth shown in Table 1 is expressed by the turbidity of the cultured liquor, which was measured by the optical density at 660 m ⁇ .
  • composition of medium is Composition of medium:

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Abstract

A microorganism of the species Lactobacillus italicus having the following taxonomic characteristics:
  • (1) Gram-positive rods in a size of 0.8-1.0 X 2-7 µ;
  • (2) formation of spore: negative;
  • (3) motility: negative;
  • (4) anaerobic to facultative aerobic;
  • (5) growth temperature: 15-38°C, optimal 28-32°C;
  • (6) growth pH: 4.2-6.5, optimal 5.4-5.8;
  • (7) catalase: negative; and
  • (8) capable of assimilating glucose and maltose to produce DL-lactic acid, ethyl alcohol and carbon dioxide but not capable of assimilating arabinose, xylose, galactose, acetic acid, citric acid, gluconic acid or propionic acid as the sole source of carbon;
in or on a sterile medium is isolated from a dough used for the preparation of panettone bread. On the basis of an unsaturated fatty acid requirement for growth, the new lactobacilli are classified into Lactobacillus italicus TL-1 (negative requirement) and Lactobacillus italicus SH150 (positive requirement). New lactobacilli are obtained by incubating an aqueous suspension of a panettone dough on an agar medium anaerobically to form colonies, from which the new lactobacilli may be selected and purely cultured. The new microorganisms are useful in the preparation of lactic acid.

Description

  • The present invention relates to microorganisms of the species Lactobacillus italicus, processes for their preparation and their use in the production of lactic acid. The lactobacilli may be isolated from a dough for making bread.
  • Lactobacillus sanfrancisco is a known microorganism isolated from a dough for sour dough French bread conventionally made in the San Francisco Bay area of the United States, and is useful in the preparation of a liquid starter and dough for sour dough bakery products such as sour dough French bread (U.S. Patents Nos. 3,734,743 and 3,891,773). This microorganism is capable of growing in the presence of a sour dough yeast, viz
  • Torulopsis holmii.
  • As a result of various studies, we have isolated . certain microorganisms from a dough used for the preparation of panettone which is a type of Italian rich bread. The present invention is based upon the discovery that the microorganisms which we have isolated are capable of growing in the absence of yeast and are of potential interest as lactic acid-producing microorganisms.
  • According to one feature of the present invention, there is provided a microorganism of the species Lactobacillus italicus having the following taxonomic characteristics:
    • (1) Gram-positive rods in a size of 0.8-1.0 x 2-7 µ;
    • (2) formation of spore: negative;
    • X3) motility: negative;
    • (4) anaerobic to facultative aerobic;
    • (5) growth temperature: 15-38°C, optimal 28-32°C;
    • (6) growth pH: 4.2-6.5, optimal 5.4-5.8;
    • (7) catalase: negative; and
    • (8) capable of assimilating glucose and maltose to produce DL-lactic acid, ethyl alcohol and carbon dioxide but not capable of assimilating arabinose, xylose, galactose, acetic acid, citric acid, gluconic acid or propionic acid as the sole source of carbon;
      in or on a sterile medium.
  • The microorganisms in one embodiment may be further characterised in that glucose and maltose are the sole assimilable carbon sources.
  • The microorganisms of the present invention have been designated by us as Lactobacillus italicus for reasons hereinafter described and include the following two strains:
    • (I) Lactobacillus italicus TL-1 (FERM-BP 189) and
    • (II) Lactobacillus italicus SH-150 (FERM-BP 190).
  • The above-mentioned two microorganisms were deposited with the Fermentation Research Institute Agency of Industrial Science and Technology, Ministry of International Trade and Industry of 1-3, Higashi 1-chome, Yatabe-machi Tsukuba-gun, Ibaraki-ken 305, Japan under the Budapest Treaty on 29th September 1982.
  • The taxonomic characteristics of these two microorganisms are identical with those of various known lactobacilli such as, for example, Lactobacillus sanfrancisco except as specified below:
  • (I) L. italicus TL-1 (FERM-BP 189): A. Morphological characteristics:
  • Where this strain is cultured in a liquid medium such as, for example, an ML medium as hereinafter defined, the present strain appears as rods having a size of (0.8-1.0) x (2 -7) µ, and occurs in the form of a single rod or in chains. This strain is non-motile, non- sporulating and Gram-positive.
  • Composition of ML medium:
    • peptone 12.5 g; yeast extract 12.5 g; potassium monohydrogenphosphate 250 mg; potassium dihydrogenphosphate 250 mg; magnesium sulfate 100 mg;
    • manganese sulfate 5.0 mg; ferrous sulfate 5.0 mg;
    • cysteine hydrochloride 300 mg; Tween 80 (commercial product of Atlas Chemical Industry, Inc., U.S.A.). 300 mg; sodium acetate 10 g; glucose 10 g;
    • maltose 10 g; water 1000 ml (adjusted to a pH of 5.6 with 1N or 6N-HC1).
    B. Cultural characteristics: (1) ML agar plate:
  • Where this strain is cultured on an ML agar plate medium prepared by adding 20 g of agar to ML medium at 30°C for 2 days, round, smooth, entire, raised, semi-transparent and milky white colonies are formed, the diameter of the colonies being not greater than about 1 mm.
  • (2) ML broth:
  • The culture exhibits a moderate turbidity. No growth on the surface layer. Flocculent sediment is formed.
  • (3) ML agar stab:
  • Growth is observed along with the stab and on the surface layer.
  • C. Physiological characteristics:
    • (1) Growth temperature: Range 15=38°C. Optimal 28-32°C.
    • (2) Growth pH: Range 4.2 to 6.5. Optimal 5.4 to 5.8.
    • (3) Relation to oxygen:
  • Anaerobic to facultative aerobic. Better growth in an atmosphere of carbon dioxide, even though growth is possible under aerobic conditions.
  • (4) Required substances for the growth:
  • An unstaturated fatty acid is not required for growth on an ML medium.
  • (5) Production of acid and gas from sugars:
  • Produced from glucose and maltose, and not produced from arabinose, xylose, ribose, fructose, mannitol, galactose, sucrose, lactose, melibiose and salicin.
  • (6) Assimilation of carbon sources:
  • Maltose and glucose are assimilable, although maltose is more strongly assimilated than glucose.
  • Arabinose, xylose, galactose, acetic acid, citric acid, gluconic acid and propionic acid are not assimilated as sole carbon sources.
    • (7) Liquefaction of gelatin: negative.
    • (8) Lithmus milk; unchanged.
    • (9) Reduction of nitrate: negative.
    • (10) Denitrification:: negative.
    • (11) Urease: negative.
    • (12) Catalase: negative.
    • (13) Oxidase: negative.
    • (14) Formation of indole: negative.
    • (15) Formation of hydrogen sulfide: negative.
    • (16) Hydrolysis of starch: negative.
    • (17) VP reaction: negative.
    • (18) DL-lactic acid and ethyl alcohol are produced from glucose or maltose at an approximate ratio of 1:0.4-0.6.
    (II) Lactobacillus italicus SH-150 (FERM-BP 190):
  • Similar taxonomic characteristics to those of L. italicus TL-1 except for the requirement of an unsaturated fatty acid for growth.
  • As hereinbefore described, L. italicus TL-1 and L. italicus SH-150 are Gram-positive rods which are non- sporulating, non-motile, anaerobic to facultative aerobic and catalase negative. Moreoever, they are capable of producing lactic acid and ethyl alcohol from glucose and maltose. Thus, it is apparent that they are classified into the genus Lactobacillus. However, the present strains differ from all lactobacilli disclosed, for example, in Bergey's Manual of Determinative Bacteriology, 8th Edition (1974) in so far as the following identifying characteristics are concerned:-
    • (a) they decompose glucose and maltose, but no other carbohydrate whilst various other known lactobacilli decompose many carbohydrates;
    • (b) the optimal growth pH is from 5.4 to 5.8, whilst various known lactobacilli exhibit an optimal pH of from 6.5 to 7.0;
    • (c) they are incapable of assimilating gluconate, whilst various other lactobacilli of the heterofermentative type are capable of decomposing gluconate;
    • (d) the present strains differ from Lactobacillus sanfrancisco which is only capable of metabolizing glucose with difficulty and produces lactic acid and acetic acid from maltose. Moreover, a fresh yeast extract is required for the growth of L. sanfrancisco.
  • For these reasons, it is believed that the microorganisms of the present invention designated by us as Lactobacillus italicus are new microorganisms.
  • The taxonomic characteristics of the present strains were determined by the methods described in "Classification and identification of microorganisms" [edited by Takeshi Hasegawa and published by Gakkai Shuppan Centre Tokyo (1975)] and Bergey's Manual of Determinative Bacteriology, 8th Edition (1974) referred to above. With regard to Lactobacillus sanfrancisco, reference was made to Applied Microbiology, 21, 459-4'65 (1971).
  • According to a further feature of the present invention there is provided a process for the preparation of a microorganism as hereinbefore defined which comprises inoculating an agar medium with an aqueous suspension of panettone dough, incubating the inoculated medium under anaerobic conditions to form colonies on said medium, isolating Lactobacillus italicus having the identifying characteristics hereinbefore defined and culturing the isolated strain(s) in or on a sterile medium.
  • Panettone dough is described in Bakers Digest October 76 80 (1977).
  • L. italicus of the present invention may be cultured, for example, in the following manner:-
  • Various solid and liquid media may be used for culturing L. italicus in so far as they are suitable for culturing lactobacilli and contain glucose and maltose as carbon sources. As carbon sources, various naturally occurring substances containing glucose and maltose such as, for example, starch hydrolyzate liquor, wheat flour and the like may.beused. If desired, various organic acids such as, for example, acetic acid, citric acid and the like may be added to the medium in order to promote the growth of L. italicus. The use of various materials containing organic nitrogen such as, for example, yeast extract, peptone, meat extract, casein, corn steep liquor, tomato juice and the like as nitrogen sources is advantageous for promoting the growth of L. italicus, and thus it is preferred to add to the medium inorganic compounds such as, for example, ammonium sulfate, ammonium nitrate, ammonium phosphate and the like. It is also advantageous to add to the medium, for example, phosphoric acid, various ions of metals such as e.g. potassium, sodium, magnesium, manganese, iron, molybdenum and the like. The addition of, for example, cysteine, cystine, glutathione, ascorbic acid and the like to the medium is also advantageous. in order to reduce the partial pressure of oxygen. Where an unsaturated fatty acid is required for the growth of the cultured microorganism, it is necessary to add a suitable agent such as, for example, oleic acid, Tween 80 (a surface active agent, commercially available from Atlas Chemical Industries, Inc., U.S.A.), lectin and the like to the medium.
  • The culturing may for example be effected at a temperature of from 15 to 38°C, preferably from 28 to 32°C and at a pH of from 4.2 - 6.5,especially from 5.4 to 5.8 for 1 to 2 days. As the present strains are more or less anaerobic, the culturing may preferably be effected under anaerobic conditions in the presence of carbon dioxide. In such a case, the mixing ratio of carbon dioxide with air may be from 25 to 95 % by volume.
  • Especially good results may be obtained, for example, by using an ML medium as hereinbefore defined in order to isolate the present strain from a dough used for the preparation of panettone as hereinbefore described. Thus, for example, a panettone dough is suspended in a sterilized physiological saline solution, suitable buffer solution and the like and the mixture is smeared, for example, on an ML agar plate medium as hereinbefore defined for culturing at a temperature of from 28 to 32°C for 1-2 days under anaerobic conditions to obtain a pure culture of L. italicus of the present invention.
  • According to a further feature of the present invention there is provided a process for the preparation of lactic acid which comprises culturing a microorganism as hereinbefore defined in or on a medium therefor whereby to obtain a cultured liquor containing lactic acid. The lactic acid obtained is preferably isolated from the obtained cultured liquor.
  • The following non-limiting Examples illustrate the present invention.
  • Example 1
  • A panettone dough purchased from the Italian market (0.3 g) was suspended in a physiological saline solution (10 ml) under. sterilized conditions. The suspension thus-prepared was multiply diluted with the same solution. Then, on each occasion, the diluted dough suspension (0.1 ml) was smeared on an ML medium as hereinbefore defined, and a Gas Pak System (Commercial Product of Baltimore Biological Laboratories, Inc., U.S.A.) was used to provide an atmosphere of carbon dioxide, in which the culturing was effected at a temperature of 30°C for 2 days to obtain semi-transparent, milky white colonies having a diameter of not greater than about 1 mm. The colonies were picked out and transferred to another ML agar plate medium for further culturing under similar conditions for 1-2 days. In this manner, 179 strains in total were isolated from 2 kinds of panettone dough, which were very similar to each other. Even though very small differences were noted between them, it was noted that all strains belonged to the same species. In view of the differences in the unsaturated fatty acid requirement for growth, Lactobacillus italicus TL-1 (negative requirement) and L. italicus SH-150 (positive requirement) were selected. In these cases, the number of the microorganisms present in the panettone doughs used was about 7 x 108 cell per gram.
  • Example 2
  • A medium containing yeast extract (3 g/l), peptone (5 g/1), KH2PO4 (0.25 g/l), MgSO4.7H2O (0.1 g/1), MnS04.4H20 (5.0 mg/1), FeS04.7H20 (5.0 mg/1), cysteine HC1 (300 mg/1) and Tween 80 (300 mg/1) and having an initial pH adjusted to 5.6 with acetic acid, was used as a basal medium, from which 6 types of media were prepared by adding to the basal medium respectively the additives containing carbon sources shown in Table 1. On each occasion, the thus-prepared medium was put into a large test tube and was sterilized at a temperature of 120°C for 15 minutes in an autoclave. Each medium was then inoculated with L. italicus TL-1 (FERM-BP 189) or L. italicus SH-150 (FERM-BP 190) (about 108 cells per ml) obtained from the ML agar plate medium hereinbefore described and incubated at a temperature of 30°C for the period shown in Table 1. The resultant growth shown in Table 1 is expressed by the turbidity of the cultured liquor, which was measured by the optical density at 660 mµ.
    Figure imgb0001
  • This table indicates that the best result was obtained by using medium No. 5.
  • Example 3
  • Preparation of lactic acid:
    • A medium having the composition stated below was inoculated with Lactobacillus italicus TL-1 (FERM-BP 189; one platinum loop) for culturing at 30°C for 18 hours to obtain a cultured liquor containing lactic acid in an amount of 0.54%:
  • Composition of medium:
    • yeast extract 12.5 g; peptone 12.5 g; glucose 11.0 g; maltose 15.0 g; KH2PO4 0.25 g; K2HPO4 0.025 g; MgSO4. 7H20 0.1 g; MgSO4.4H2O 5 mg; FeS04.7H20 5 mg; Tween 80 300 mg; L-cysteine HC1 300 mg; adenine 20 mg; guanine 20 mg; uracil 20 mg; sodium acetate. 3H20 10.0 g (made up to 1000 ml with water; pH being adjusted to 5.6 with hydrochloric acid).

Claims (10)

1. A microorganism of the species Lactobacillus italicus having the following taxonomic characteristics:
(1) Gram-positive rods in a size of 0.8-1.0 x 2-7 µ;
(2) formation of spore: negative;
(3) motility: negative;
(4) anaerobic to faculative aerobic;
(5) growth temperature: 15-38°C. optimal 28-32°C;
(6) growth pH: 4.2-6.5, optimal 5.4-5.8;
(7) catalase: negative; and
(8) capable of assimilating glucose and maltose to produce DL-lactic acid, ethyl alcohol and carbon dioxide but not capable of assimilating arabinose, xylose, galactose, acetic acid, citric acid, gluconic acid or propionic acid as the sole source of carbon;
in or on a sterile medium.
2. A microorganism as claimed in claim 1 wherein glucose and maltose are the sole assimilable carbon sources.
3. A microorganism as claimed in claim 1 having no unsaturated fatty acid requirement for growth, said microorganism being designated Lactobacillus italicus TL-1 (FERM-BP 189).
4. A microorganism as claimed in claim 1 having an unsaturated fatty acid requirement for growth, said microorganism being designated Lactobacillus italicus SH 150 (FERM-BP 190)
5. A process for preparing a microorganism as claimed in claim 1 which comprises inoculating an agar medium with an aqueous suspension of a panettone dough, incubating the inoculated medium under anaerobic conditions to form colonies on said medium, isolating Lactobacillus italicus having the identifying characteristics defined in claim 1 and culturing the isolated strain(s) in or on a sterile medium.
6. A process as claimed in claim 5 wherein the isolation of Lactobacillus italicus comprises the isolation of' Lactobacillus italicus TL-1 (FERM-BP 189) which has no unsaturated fatty acid requirement for growth and Lactobacillus italicus SH 150 (FERM-BP 190) which has an unsaturated fatty acid requirement for growth.
7. A process as claimed in claim 5 or claim 6 wherein the incubation is effected at a temperature of from 15-38°C for 1-2 days.
8. A process as claimed in any one of claims 5 to 7 wherein the incubation is effected at a pH of from 4.2 - 6.5.
9. A process for the preparation of lactic acid which comprises culturing a microorganism as claimed in claim 1 in or on a medium therefor whereby to obtain a cultured liquor containing lactic acid.
10. A process as claimed in claim 9 wherein the lactic acid is isolated from the obtained cultured liquor.
EP83307542A 1982-12-11 1983-12-12 Microorganisms of the species lactobacillus italicus, processes for their preparation and their use in the production of lactic acid Expired EP0113215B1 (en)

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Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4758345A (en) * 1986-02-28 1988-07-19 The United States Of America As Represented By The United States Department Of Energy Anaerobic microbial dissolution of lead and production of organic acids
WO1995019425A1 (en) * 1994-01-18 1995-07-20 Institut Français De Recherche Scientifique Pour Le Developpement En Cooperation (O.R.S.T.O.M.) Bacterial strains of genus bacillus closely phenotypically related to genus lactobacillus, culture method and use
US5516684A (en) * 1994-03-11 1996-05-14 The Calpis Food Industry Co., Ltd. Biologically pure culture of Lactobacillus acidophilus FERM-P-14204 or FERM-P-14205
US5541111A (en) * 1993-11-04 1996-07-30 The Calpis Food Industry Co., Ltd. Lactobacillus helveticus mutants having low increase in acidity of lactic acid during storage

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS61181333A (en) * 1985-02-06 1986-08-14 敷島製パン株式会社 Dough modifier

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GB936339A (en) * 1961-06-20 1963-09-11 Melle Usines Sa Process for extracting carboxylic acids produced by fermentation
FR2126452A1 (en) * 1971-02-26 1972-10-06 Leo Kline
WO1980002282A1 (en) * 1979-04-20 1980-10-30 B Bodenrader Method for bacteriological treatment of manure and high bod industrial wastes

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GB936339A (en) * 1961-06-20 1963-09-11 Melle Usines Sa Process for extracting carboxylic acids produced by fermentation
FR2126452A1 (en) * 1971-02-26 1972-10-06 Leo Kline
WO1980002282A1 (en) * 1979-04-20 1980-10-30 B Bodenrader Method for bacteriological treatment of manure and high bod industrial wastes

Cited By (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4758345A (en) * 1986-02-28 1988-07-19 The United States Of America As Represented By The United States Department Of Energy Anaerobic microbial dissolution of lead and production of organic acids
US5541111A (en) * 1993-11-04 1996-07-30 The Calpis Food Industry Co., Ltd. Lactobacillus helveticus mutants having low increase in acidity of lactic acid during storage
US5695796A (en) * 1993-11-04 1997-12-09 The Calpis Food Industry Co., Ltd. Fermented milk product
WO1995019425A1 (en) * 1994-01-18 1995-07-20 Institut Français De Recherche Scientifique Pour Le Developpement En Cooperation (O.R.S.T.O.M.) Bacterial strains of genus bacillus closely phenotypically related to genus lactobacillus, culture method and use
FR2715166A1 (en) * 1994-01-18 1995-07-21 Orstom Bacterial strains phylogenetically close to the genus Lactobacillus and the genus Bacillus, culture method and applications.
US6022537A (en) * 1994-01-18 2000-02-08 Institut Francais De Recherche Scientifique Pour Le Developpement En Cooperation (Orstom) Bacterial strains of the genus Bacillus, closely phenotypically related to the genus Lactobacillus, culture method and use
US5516684A (en) * 1994-03-11 1996-05-14 The Calpis Food Industry Co., Ltd. Biologically pure culture of Lactobacillus acidophilus FERM-P-14204 or FERM-P-14205
US5707854A (en) * 1994-03-11 1998-01-13 Calpis Food Industry Co., Ltd. Lactic acid bacteria of the Genus lactobacillus

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EP0113215B1 (en) 1988-09-21
EP0113215A3 (en) 1986-06-11
JPH0145352B2 (en) 1989-10-03
DE3378057D1 (en) 1988-10-27
JPS59106291A (en) 1984-06-19

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